EP0018628A1 - Method of determining gamma-glutamyl transpeptidase - Google Patents

Method of determining gamma-glutamyl transpeptidase Download PDF

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Publication number
EP0018628A1
EP0018628A1 EP80102321A EP80102321A EP0018628A1 EP 0018628 A1 EP0018628 A1 EP 0018628A1 EP 80102321 A EP80102321 A EP 80102321A EP 80102321 A EP80102321 A EP 80102321A EP 0018628 A1 EP0018628 A1 EP 0018628A1
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Prior art keywords
acid
glutamyl
nitranilide
substrate
solution
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EP80102321A
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German (de)
French (fr)
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EP0018628B1 (en
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Helmut Dr. Heber
Helmut Dr. Kohl
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Siemens Healthcare Diagnostics GmbH Germany
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Behringwerke AG
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/48—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving transferase

Definitions

  • the invention relates to a substrate for determining the ⁇ -glutamyl transpeptidase in aqueous systems, especially in biological liquids, in particular in blood serum. It is mainly used for the detection and diagnosis of liver diseases.
  • the release of the yellow-colored p-nitraniline can be followed optically. It is a direct measure of the existing y-GT activity.
  • nitraniline is e.g.
  • aqueous solutions with pH values such as must be observed when determining the ⁇ -GT.
  • This is known, inter alia, from patent applications DE-OS 20 42 829 and DE-OS 22 59 512.
  • the substrate was pre-dissolved in dilute hydrochloric acid or in organic solvents in high concentration and added to the determination batches in this form, thereby avoiding a sharp shift in the pH.
  • the disadvantage of this less elegant method is the instability of the substrate, which is observed in a strongly acidic, aqueous environment. Therefore, such solutions can only be used for measurements at short notice.
  • the y-glutamyl-p-nitranilide is dissolved at a temperature between 50 and 60 ° C and then cooled to room temperature.
  • a risk of spontaneous hydrolysis of the chromophoric group is a risk of spontaneous hydrolysis of the chromophoric group.
  • the dissolution process is time consuming and often not successful anyway because of the recrystallization of the substrate.
  • the substrate y-glutamyl-p-nitroanilide should be sufficiently soluble in the applicable for the implementation of the determination temperature of about 25 0 C.
  • y-glutamyl-p-nitranilide preferably as a lyophilisate, dissolves in the test buffer at 20 to 25 ° C. almost immediately when the substrate is protonized, that is to say as a salt.
  • the y-glutamyl-p-nitranilide is mixed with an acid in a stoichiometric ratio or in excess of the base equivalents before drying.
  • the invention accordingly relates to a dry preparation of the ⁇ -glutamyl-p-nitranilide for use as a substrate for determining the y-GT, characterized by the content of an acid in at least a molar ratio to its base equivalent.
  • the reagent has better color stability even after redissolution, if it is mixed with a quaternary polycation, it is part of the subject of the invention that the dry preparation is optionally mixed with such a high molecular weight solution stabilizer.
  • ammonium salts of ⁇ -glutamyl-p-nitranilide are optionally used with an excess of acid as the substrate for the determination of y-glutamyl transpeptidase
  • organic or inorganic acids capable of forming an ammonium salt of y-glutamyl-p-nitranilide, e.g. Sulfuric acid, the phosphoric acids, all hydrogen halide acids, oxyhalogenide acids, acetic acid and trifluoroacetic acid.
  • Combinations of an inorganic and an organic acid such as di-, tri-, tetra-carboxylic acids or -hydroxycarboxylic acids, for example malonic acid, succinic acid, glutaric acid, adipic acid, pimelic acid, maleic acid, fumaric acid, malic acids, tartaric acids, citric acid, ascorbic acid, glucuronic acid, have proven successful.
  • an inorganic acid e.g. Hydrochloric acid
  • an organic acid e.g. Tartaric acid used.
  • Relative to one base equivalent of the nitranilide, the ratio of about 0.9 equivalents of the inorganic acid and about 0.1 equivalents of the organic acid is particularly favorable.
  • Another very simple way of producing the preparation according to the invention is to acidify the aqueous solution of the nitranilide with the desired acid, after which the drying can be carried out.
  • the main advantage of the dry! product is that there is no significant hydrolysis before it is used in the test, and accordingly there is no disturbance due to a blank value of high optical extinction.
  • the substrate property is improved by adding a quaternary polycation to the substrate in a concentration of 0.1-100 mg / ml test solution.
  • the salt of Y-glutamyl-p-nitranilide, optionally mixed with the polymer, is kept in solid form. This can be done in dry fillings or compressed in tablets.
  • mannitol is used in a concentration range of 1-50 mg / ml.
  • the glycylglycine involved in the reaction of the transpeptidase can also already be present in the dry preparation, so that for the test only the substrate mixture has to be dissolved in water, after which the enzyme can be determined.

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Abstract

1. Process for quantitatively determining gamma-glutamyltranspeptidase by means of a lyophilized salt of gamma-glutamyl-p-nitranilide with an acid, which process comprises dissolving that salt in a buffer, adding the sample containing the gamma-glutamyltranspeptidase and determining the amount of p-nitraniline set free.

Description

Die Erfindung betrifft ein Substrat zur Bestimmung der y-Glutamyltranspeptidase in wässrigen Systemen, vor allem in biologischen Flüssigkeiten, insbesondere in Blutserum. Es findet hauptsächlich Verwendung zur Erkennung und Verlaufsdiagnose von Lebererkrankungen.The invention relates to a substrate for determining the γ-glutamyl transpeptidase in aqueous systems, especially in biological liquids, in particular in blood serum. It is mainly used for the detection and diagnosis of liver diseases.

Die Bestimmung der y-Glutamyltranspeptidase (y-GT) - Aktivität erfolgt nach dem bekannten Reaktionsschema:

  • γ-Glutamyl-p-Nitranilid + Glycylglycin⇄p-Nitranilin + Glutamylglycylglycin.
The determination of the y-glutamyl transpeptidase (y-GT) activity is carried out according to the known reaction scheme:
  • γ-glutamyl-p-nitranilide + glycylglycine⇄p-nitraniline + glutamylglycylglycine.

Die Freisetzung des gelbgefärbten p-Nitranilins kann optisch verfolgt werden. Sie ist ein direktes Maß für die vorhandene y-GT-Aktivität.The release of the yellow-colored p-nitraniline can be followed optically. It is a direct measure of the existing y-GT activity.

Der Vorzug des einfachen Nachweises des Nitranilins ist z. Zt. mit dem Nachteil verknüpft, daß das Substrat y-Glutamyl-p-Nitranilid in wässrigen Lösungen mit pH-Werten,wie sie bei der Bestimmung der γ-GT eingehalten werden müssen, sehr schlecht löslich ist. Dies ist u.a. aus den Patentanmeldungen DE-OS 20 42 829 und DE-OS 22 59 512 bekannt. Man hat schon mehrfach versucht, dieses Problem zu lösen. So wurde das Substrat beispielsweise in verdünnter Salzsäure oder in organischen Lösungsmitteln in hoher Konzentration vorgelöst und in dieser Form den Bestimmungsansätzen zugefügt, wodurch eine starke Verschiebung des pH-Wertes vermieden wird. Zu dem Nachteil dieser wenig eleganten Methode kommt noch die Instabilität des Substrates hinzu, die in stark saurem, wässrigen Milieu beobachtet wird. Daher sind solche Lösungen nur kurzfristig für Messungen einsetzbar.The advantage of simple detection of nitraniline is e.g. Currently associated with the disadvantage that the substrate y-glutamyl-p-nitranilide is very poorly soluble in aqueous solutions with pH values, such as must be observed when determining the γ-GT. This is known, inter alia, from patent applications DE-OS 20 42 829 and DE-OS 22 59 512. Several attempts have been made to solve this problem. For example, the substrate was pre-dissolved in dilute hydrochloric acid or in organic solvents in high concentration and added to the determination batches in this form, thereby avoiding a sharp shift in the pH. The disadvantage of this less elegant method is the instability of the substrate, which is observed in a strongly acidic, aqueous environment. Therefore, such solutions can only be used for measurements at short notice.

Gemäß einem anderen bekannten Verfahren wird das y-Glutamyl-p-Nitranilid bei einer Temperatur zwischen 50 und 60°C aufgelöst und danach auf Zimmertemperatur abgekühlt. Auch hierbei besteht die Gefahr der spontanen Hydrolyse der chromophoren Gruppe. Weiterhin ist das Auflöseverfahren zeitraubend und häufig wegen der Rekristallisation des Substrates ohnehin nicht erfolgreich.According to another known method, the y-glutamyl-p-nitranilide is dissolved at a temperature between 50 and 60 ° C and then cooled to room temperature. Here too there is a risk of spontaneous hydrolysis of the chromophoric group. Furthermore, the dissolution process is time consuming and often not successful anyway because of the recrystallization of the substrate.

Solche Methoden sind für die routinemäßige Anwendung des Testes in klinischen Laboratorien unbefriedigend, so daß die Aufgabe bestand, die damit verbundenen Nachteile zu beseitigen. Das Substrat y-Glutamyl-p-Nitranilid soll bei der für die Durchführung der Bestimmung einzuhaltenden Temperatur von etwa 250C ausreichend gut löslich sein.Such methods are unsatisfactory for the routine use of the test in clinical laboratories, so that the task was to eliminate the associated disadvantages. The substrate y-glutamyl-p-nitroanilide should be sufficiently soluble in the applicable for the implementation of the determination temperature of about 25 0 C.

Erfindungsgemäß wurde nun gefunden, daß sich trockenes y-Glutamyl-p-Nitranilid, vorzugsweise als Lyophilisat, im Testpuffer bei 20 bis 25°C nahezu augenblicklich auflöst, wenn das Substrat protonisiert, das heißt als Salz vorliegt. Dies bedeutet, daß das y-Glutamyl-p-Nitranilid vor der Trocknung mit einer Säure im stöchiometrischen Verhältnis oder im Überschuß zu den Basenäquivalenten versetzt wird.According to the invention, it has now been found that dry y-glutamyl-p-nitranilide, preferably as a lyophilisate, dissolves in the test buffer at 20 to 25 ° C. almost immediately when the substrate is protonized, that is to say as a salt. This means that the y-glutamyl-p-nitranilide is mixed with an acid in a stoichiometric ratio or in excess of the base equivalents before drying.

Gegenstand der Erfindung ist demnach eine trockene Zu- bereitung des γ-Glutamyl-p-Nitranilids zur Verwendung als Substrat zur Bestimmung der y-GT, gekennzeichnet durch den Gehalt einer Säure in mindestens molarem Verhältnis zu dessen Basenäquivalent.The invention accordingly relates to a dry preparation of the γ-glutamyl-p-nitranilide for use as a substrate for determining the y-GT, characterized by the content of an acid in at least a molar ratio to its base equivalent.

Da.zudem gefunden wurde, daß das Reagenz auch nach Wiederauflösung eine bessere Farbstabilität besitzt, wenn es mit einem quaternären Polykation versetzt wird, gehört es zum Gegenstand der Erfindung, daß die trockene Zubereitung gegebenenfalls mit einem solchen hochmolekularen Lösungsstabilisator versetzt ist.Since it has also been found that the reagent has better color stability even after redissolution, if it is mixed with a quaternary polycation, it is part of the subject of the invention that the dry preparation is optionally mixed with such a high molecular weight solution stabilizer.

Im engeren Sinne gehört es zum Gegenstand der Erfindung, daß als Substrat für die Bestimmung von y-Glutamyltranspeptidase die Ammoniumsalze des γ-Glutamyl-p-Nitranilids gegebenenfalls mit einem Überschuß an Säure verwendet werdenIn the narrower sense, it is part of the subject matter of the invention that the ammonium salts of γ-glutamyl-p-nitranilide are optionally used with an excess of acid as the substrate for the determination of y-glutamyl transpeptidase

Als Säuren, die hierfür in Frage kommen, sind praktisch alle organischen oder anorganischen Säuren zu nennen, die ein Ammoniumsalz des y-Glutamyl-p-Nitranilids zu bilden vermögen, z.B. Schwefelsäure, die Phosphorsäuren, alle Halogenwasserstoff-Säuren, Oxyhalogenwasserstoff-Säuren, Essigsäure und Trifluoressigsäure. Bewährt haben sich Kombinationen aus einer anorganischen und einer organischen Säure wie Di-, Tri-, Tetra-Carbonsäuren beziehungsweise -Hydroxycarbonsäuren, beispielsweise Malonsäure, Bernsteinsäure, Glutarsäure, Adipinsäure, Pimelinsäure, Maleinsäure, Fumarsäure, Äpfelsäuren, Weinsäuren, Citronensäure, Ascorbinsäure, Glucuronsäure. Vorzugsweise wird eine anorganische Säure, z.B. Salzsäure, in Kombination mit einer organischen Säure, z.B. Weinsäure, verwendet. Bezogen auf ein Basenäquivalent des Nitranilids ist das Verhältnis von etwa 0,9 Äquivalenten der anorganischen Säure und etwa 0,1 Äquivalenten der organischen Säure besonders günstig.Practically all organic or inorganic acids capable of forming an ammonium salt of y-glutamyl-p-nitranilide, e.g. Sulfuric acid, the phosphoric acids, all hydrogen halide acids, oxyhalogenide acids, acetic acid and trifluoroacetic acid. Combinations of an inorganic and an organic acid such as di-, tri-, tetra-carboxylic acids or -hydroxycarboxylic acids, for example malonic acid, succinic acid, glutaric acid, adipic acid, pimelic acid, maleic acid, fumaric acid, malic acids, tartaric acids, citric acid, ascorbic acid, glucuronic acid, have proven successful. Preferably an inorganic acid, e.g. Hydrochloric acid, in combination with an organic acid, e.g. Tartaric acid used. Relative to one base equivalent of the nitranilide, the ratio of about 0.9 equivalents of the inorganic acid and about 0.1 equivalents of the organic acid is particularly favorable.

Zur Herstellung des Substrates eignen sich alle gängigen Methoden der organischen Chemie zur Herstellung von Ammoniumsalzen von primären Aminen. Ein Überschuß der Säuren ist so lange nicht schädlich, so lange das Substrat ohne Nachteile gefriergetrocknet werden kann und das wiederaufgelöste Substrat die Kapazität des Testpuffers nicht überschreitet. Am einfachsten geht man von einer wässrigen Lösung des Y-Glutamyl-p-Nitranilids aus, stellt das stöchiometrische Verhältnis des Anilids zu einer Säure fest, versetzt die Lösung mit der betreffenden Säure in dem gewünschten Verhältnis, gegebenenfalls mit einem Überschuß von ca. 20 % und trocknet das Produkt, vorzugsweise durch Gefriertrocknung.All common methods of organic chemistry are suitable for producing the substrate Ammonium salts of primary amines. An excess of the acids is not harmful as long as the substrate can be freeze-dried without disadvantages and the redissolved substrate does not exceed the capacity of the test buffer. It is easiest to start with an aqueous solution of the Y- glutamyl-p-nitranilide, determine the stoichiometric ratio of the anilide to an acid, add the desired acid to the solution, optionally with an excess of about 20% and dries the product, preferably by freeze drying.

Ein anderer sehr einfacher Weg zur Herstellung der er- findungsgemäßen Zubereitung ist, die wässrige Lösung des Nitranilids mit der gewünschten Säure anzusäuern, wonach die Trocknung durchgeführt werden kann.Another very simple way of producing the preparation according to the invention is to acidify the aqueous solution of the nitranilide with the desired acid, after which the drying can be carried out.

Der wesentliche Vorteil des erfindungsgemäßen Trocken- ! produktes besteht darin, daß es hierbei zu keiner nennenswerten Hydrolyse vor dessen Verwendung im Test kommt und demgemäß eine Störung durch einen Leerwert hoher opti- scher Extinktion ausbleibt.The main advantage of the dry! product is that there is no significant hydrolysis before it is used in the test, and accordingly there is no disturbance due to a blank value of high optical extinction.

Eine Verbesserung der Substrateigenschaft wird, wie bereits erwähnt, dadurch erreicht, daß dem Substrat ein quaternäres Polykation in einer Konzentration von 0,1 - 100 mg/ml Testlösung zugesetzt wird. Dieser Befund ist überraschend, da es bekannt ist, daß übliche kationische bzw. anionische Detergenzien die y-Glutamyltranspeptidase Aktivität hemmen bzw. vermindern. Demgegenüber beeinflus sen quaternäre Polykationen auf Basis von Polyvinylpyrrolidon die Enzymaktivität nicht meßbar. Den gleicher Effekt zeigen äthoxylierte Alkylphenole oder äthoxyliertE Phenole sowie Tetrafluoräthylen-Polymerisate mit verzweigten Perfluorgruppen. Diese Polymeren verzögern einzeln oder in Kombination die Rekristallisation des γ-Glutamyl-p-Nitranilids bei Raumtemperatur.As already mentioned, the substrate property is improved by adding a quaternary polycation to the substrate in a concentration of 0.1-100 mg / ml test solution. This finding is surprising since it is known that customary cationic or anionic detergents inhibit or reduce the y-glutamyl transpeptidase activity. In contrast, quaternary polycations based on polyvinylpyrrolidone have no measurable influence on enzyme activity. The same effect is shown by ethoxylated alkylphenols or ethoxylated phenols and tetrafluoroethylene polymers with branched perfluoro groups. These polymers delay individually or in combination the recrystallization of the γ-glutamyl-p-nitranilide at room temperature.

Das gegebenenfalls mit dem Polymeren versetzte Salz des Y-Glutamyl-p-Nitranilids wird in fester Form zur Verfügung gehalten. Dies kann in Trockenabfüllungen oder in Tabletten verpreßt geschehen.The salt of Y-glutamyl-p-nitranilide, optionally mixed with the polymer, is kept in solid form. This can be done in dry fillings or compressed in tablets.

Weitere Verbesserungen hinsichtlich der Stabilität des Produktes werden erreicht durch übliche Zuschläge bei der Gefriertrocknung von Produkten wie Gelatine oder Gelatine-Hydrolysaten, Zuckeralkoholen oder Kohlenhydraten, Pufferkomponenten, Konservierungsmitteln und/oder Lösungsvermittlern. In der bevorzugten Form wird Mannit verwendet in einem Konzentrationsbereich von 1 - 50 mg/ml.Further improvements with regard to the stability of the product are achieved by customary supplements in the freeze drying of products such as gelatin or gelatin hydrolyzates, sugar alcohols or carbohydrates, buffer components, preservatives and / or solubilizers. In the preferred form, mannitol is used in a concentration range of 1-50 mg / ml.

Auch das in die Reaktion der Transpeptidase eingehende Glycylglycin kann in der trockenen Zubereitung bereits enthalten sein, so daß für den Test lediglich das Substratgemisch in Wasser aufgelöst werden muß, wonach das Enzym bestimmt werden kann. ,The glycylglycine involved in the reaction of the transpeptidase can also already be present in the dry preparation, so that for the test only the substrate mixture has to be dissolved in water, after which the enzyme can be determined. ,

Das folgende Beispiel soll die Erfindung näher erläutern.The following example is intended to explain the invention in more detail.

BeipielExample

Das erfindungsgemäße Substrat wird wie folgt hergestellt:

  • a) Substratkomponenten
    • 1) 1,17 g γ-Glutamyl-p-nitranilid (1,176 g/1000 ml = 4,4 mM)
    • 2) 8,7 g Glycylglycin
    • 3) 16 g Mannit
    • 4) 10 g Kalium-Natrium Tartrat . 4 H20
    • 5) 40 g Quarternäres Polyamin
    • 6) 2 n HCl bis pH 2-3
    • 7) dest. Wasser → 1 Liter
    Die Komponenten 1 - 5 werden in ca. 600 bis 700 ml dest. Wasser aufgenommen, auf etwa 40°C erwärmt und mit der Komponenten 6 bis zu einem pH-Wert zwischen 3 und 4 versetzt. Nach vollständiger Auflösung aller Substratkomponenten wird die Lösung auf etwa 20°C abgekühlt und der pH-Wert mit 2 n HCl (6) auf 2 bis 3 eingestellt. Die Lösung wird schließlich mit destilliertem Wasser (7) auf 1,0 Liter aufgefüllt. Es hat sich als zweckmäßig erwiesen, die Bestimmung der γ-Glutamyltranspeptidase mit 2 ml dieser Substratlösung durchzuführen, so daß man hierfür Abfüllungen von 2 ml herstellt und anschließend gefriertrocknet Für die Durchführung des Testes wird das gefriergetrocknete Produkt mit der Pufferlösung b) wiederaufgelöst.
  • b) Pufferlösung
    • 8) 24 g Tris-(hydroxymethyl)-aminomethan
    • 9) 5 g Natriumazid
    werden in etwa 800 ml dest. Wassers gelöst, mit 2 n HCl auf pH 9,2 bis 9,5 eingestellt und mit dest. Wasser auf 1,0 Liter aufgefüllt. Wird das lyophilisierte Substratgemisch nach a) mit 2 ml Pufferlösunq der erwähnten Zusammensetzung,versetzt, stellt sich ein pH-Wert von etwa 8,2 ein.
The substrate according to the invention is produced as follows:
  • a) Substrate components
    • 1) 1.17 g γ-glutamyl-p-nitranilide (1.176 g / 1000 ml = 4.4 mM)
    • 2) 8.7 g glycylglycine
    • 3) 16 g mannitol
    • 4) 10 g of potassium sodium tartrate. 4 H 2 0
    • 5) 40 g quaternary polyamine
    • 6) 2N HCl to pH 2-3
    • 7) least Water → 1 liter
    Components 1 - 5 are distilled in approx. 600 to 700 ml. Water taken up, heated to about 40 ° C and mixed with component 6 to a pH between 3 and 4. After all substrate components have completely dissolved, the solution is cooled to about 20 ° C. and the pH is adjusted to 2 to 3 with 2N HCl (6). The solution is finally made up to 1.0 liter with distilled water (7). It has proven to be expedient to carry out the determination of the γ-glutamyl transpeptidase with 2 ml of this substrate solution, so that bottlings of 2 ml are prepared for this purpose and then freeze-dried. To carry out the test, the freeze-dried product is redissolved with the buffer solution b).
  • b) buffer solution
    • 8) 24 g tris (hydroxymethyl) aminomethane
    • 9) 5 g sodium azide
    are distilled in about 800 ml. Dissolved water, adjusted to pH 9.2 to 9.5 with 2N HCl and with dist. Make up to 1.0 liters of water. If 2 ml of buffer solution of the composition mentioned is added to the lyophilized substrate mixture according to a), a pH of about 8.2 is established.

Claims (3)

1. Trockene Zubereitung des y-Glutamyl-Nitranilids zur Verwendung als Substrat zur Bestimmung der Y-Glutamyltranspeptidase, gekennzeichnet dadurch, daß sie eine Säure in mindestens molarem Verhältnis zum Basenäquivalent des Nitranilids und gegebenenfalls einen Lösungsstabilisator enthält.1. Dry preparation of the y-glutamyl nitranilide for use as a substrate for determining the Y glutamyl transpeptidase, characterized in that it contains an acid in at least a molar ratio to the base equivalent of the nitranilide and optionally a solution stabilizer. 2. Zubereitung gemäß Anspruch 1, dadurch gekennzeichnet, daß der Lösungsstabilisator ein quaternäres Polykation auf der Basis von Polyvinylpyrrolidon, einem äthoxylierten Alkylphenol, einem äthoxylierten Phenol oder einem Tetrafluoräthylen-Polymerisat mit verzweigten Perfluorgruppen ist.2. Preparation according to claim 1, characterized in that the solution stabilizer is a quaternary polycation based on polyvinylpyrrolidone, an ethoxylated alkylphenol, an ethoxylated phenol or a tetrafluoroethylene polymer with branched perfluoro groups. 3. Zubereitung gemäß Anspruch 1, dadurch gekennzeichnet, daß der Lösungsstabilisator in einer Konzentration von 0,1 - 100 mg/ml Testlösung vorliegt.3. Preparation according to claim 1, characterized in that the solution stabilizer is present in a concentration of 0.1-100 mg / ml test solution.
EP80102321A 1979-05-04 1980-04-30 Method of determining gamma-glutamyl transpeptidase Expired EP0018628B1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AT80102321T ATE7049T1 (en) 1979-05-04 1980-04-30 PROCEDURE FOR DETERMINING GAMMAGLUTAMYL TRANSPEPTIDASE.

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DE19792917999 DE2917999A1 (en) 1979-05-04 1979-05-04 SUBSTRATE FOR DETERMINING THE GAMA GLUTAMYL TRANSPEPTIDASE
DE2917999 1979-05-04

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EP0018628A1 true EP0018628A1 (en) 1980-11-12
EP0018628B1 EP0018628B1 (en) 1984-04-11

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JP (1) JPS55153600A (en)
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CA (1) CA1148455A (en)
DE (2) DE2917999A1 (en)
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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1984000978A1 (en) * 1982-08-30 1984-03-15 Beckman Instruments Inc METHOD FOR DETERMINING gamma-GLUTAMYLTRANSFERASE ACTIVITY AND KITS CONTAINING A NOVEL SUBSTRATE SOLUTION FOR USE THEREIN
WO1984004931A1 (en) * 1983-06-08 1984-12-20 Coulter Electronics Method and composition for determination of gamma glutamyl transpeptidase
US4511651A (en) * 1982-07-30 1985-04-16 American Monitor Corporation Reagent composition and assay for the determination of γ-glutamyltransferase activity
EP0266905A1 (en) * 1986-10-07 1988-05-11 Unitika Ltd. Reagents for assay of gamma-glutamyl-transpeptidase

Families Citing this family (2)

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JPS53147034A (en) * 1977-05-30 1978-12-21 Wako Pure Chem Ind Ltd Gamma-glutamyl-p-aminoanilide derivatives and process for their preparation
US4588836A (en) * 1982-09-01 1986-05-13 Toyo Jozo Kabushiki Kaisha Novel synthetic substrate and assay method using the same

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Cited By (5)

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Publication number Priority date Publication date Assignee Title
US4511651A (en) * 1982-07-30 1985-04-16 American Monitor Corporation Reagent composition and assay for the determination of γ-glutamyltransferase activity
WO1984000978A1 (en) * 1982-08-30 1984-03-15 Beckman Instruments Inc METHOD FOR DETERMINING gamma-GLUTAMYLTRANSFERASE ACTIVITY AND KITS CONTAINING A NOVEL SUBSTRATE SOLUTION FOR USE THEREIN
WO1984004931A1 (en) * 1983-06-08 1984-12-20 Coulter Electronics Method and composition for determination of gamma glutamyl transpeptidase
US4560650A (en) * 1983-06-08 1985-12-24 Coulter Electronics, Inc. Method and compositions for determination of gamma glutamyl transpeptidase
EP0266905A1 (en) * 1986-10-07 1988-05-11 Unitika Ltd. Reagents for assay of gamma-glutamyl-transpeptidase

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ES490961A0 (en) 1980-12-01
CA1148455A (en) 1983-06-21
AU5803280A (en) 1980-11-06
AU544691B2 (en) 1985-06-13
ES8102358A1 (en) 1980-12-01
DE3067415D1 (en) 1984-05-17
JPS55153600A (en) 1980-11-29
EP0018628B1 (en) 1984-04-11
ATE7049T1 (en) 1984-04-15
DE2917999A1 (en) 1980-11-13

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